黄素单核苷酸对羟自由基损伤的大鼠肝线粒体复合物Ⅰ的保护作用

目的:观察黄素单核苷酸(FMN) 对羟自由基(OH·) 损伤线粒体酶复合物1(complex Ⅰ) 的作用,分析FMN与complex Ⅰ的相互作用机制.方法:差速离心法分离大鼠肝线粒体,OH·体外损伤线粒体,氧电极法测定线粒体耗氧速率,比色法检测线粒体complex Ⅰ动力学参数及线粒体内MDA、GSH 含量及SOD活性.结果:OH·损伤后,线粒体3态、4态(state3,state4) 耗氧速率、complex Ⅰ最大反应速度(Vm) 降低(P<0.05) ,但呼吸控制率(respiratory control ratio,RCR) 、磷氧比(P/O) 及complex Ⅰ米氏常数(Km) 不变,线粒体中GSH含量下降(P<0.05) ,SOD活性降低(P<0.05) ,MDA含量升高.损伤线粒体补充FMN 165 ng/ml 可部分恢复complex Ⅰ呼吸途径state3耗氧速率(P<0.05) ,补充FMN 82.5～165 ng/mg可显著提高OH·损伤的线粒体复合物Ⅰ的Vm(P<0.05) .损伤线粒体补充较低剂量的FMN(FMN<82.5 ng FMN/ml),线粒体中GSH、SOD、MDA含量不发生变化,在补充较高剂量FMN(>165 ng FMN/ml)时 ,SOD活性升高(P<0.05) ,但线粒体MDA含量升高(P<0.05) 、GSH含量下降(P<0.05) .结论:FMN能够提高氧化损伤线粒体复合物Ⅰ活性,促进complex Ⅰ呼吸途径的state3耗氧速率.FMN不能直接清除OH·及自由基,可能通过保护complex Ⅰ中的酶结合/活性位点免受OH·等自由基攻击从而维持酶活性.     

体外评价口服降糖药对HepG2细胞线粒体功能的影响

目的:评价DPP- Ⅳ抑制剂类口服降糖药A化合物的线粒体毒性,从而探讨A化合物的可能毒性机制?方法:HepG2细胞培养基中分别加入曲格列酮50~300 μmol/L,培养24 h,或加入A化合物溶液100~300 μmol/L,培养24 h,CCK8细胞计数法测定细胞存活率?HepG2细胞培养基中分别加入曲格列酮100?200和225 μmol/L培养24 h,或加入A化合物溶液100?150和200 μmol/L培养24 h,化学发光法检测胞内ATP合成水平,荧光探针法检测胞内钙离子浓度?活性氧?线粒体膜电位(?驻Ψm)变化,透射电镜观察线粒体超微结构?结果:与溶媒对照组相比,曲格列酮50~300 μmol/L可以抑制细胞存活率,IC50为178 μmol/L;A化合物 100~300 μmol/L对细胞存活率有抑制作用,IC50为159 μmol/L?与溶媒对照组相比,曲格列酮≥200 μmol/L时导致线粒体ATP显著下降(P<0.01)??驻Ψm显著性下降(P < 0.01),≥100 μmol/L时可致活性氧水平和钙离子浓度显著升高(P < 0.05或P < 0.01)?A化合物≥100 μmol/L时ATP合成水平显著降低?钙离子浓度显著升高(P < 0.01),≥150 μmol/L时活性氧水平明显升高(P < 0.01),≥200 μmol/L时?驻Ψm显著性下降(P < 0.05),并可导致线粒体结构发生病变?结论:DPP- Ⅳ抑制剂类口服降糖药A化合物可以通过干扰线粒体代谢功能和结构而诱导线粒体损伤?     

线粒体源性氧自由基参与二氮嗪预处理的心肌保护作用

目的：研究线粒体ATP敏感性钾通道开放剂二氮嗪预处理中氧自由基（ROS）的来源。方法：传代培养的未成熟SD大鼠心肌细胞，随机分为对照组、二氮嗪（30μmol/L)预处理组、二联苯碘（DPI，10μmol/L)处理组、myxothiazol(0.2μmol/L)处理组。实验结束时全自动生化仪检测培养液中乳酸脱氢酶（LDH）活性，硫代巴比妥酸法测细胞内丙二醛（MDA）含量，台盼蓝染色法统计死亡率，电镜下观察线粒体的形态。结果：二氮嗪预处理可降低细胞死亡率、细胞内MDA含量及培养液中LDH的释放量，减轻细胞内线粒体水肿程度。二氮嗪的这种保护作用可以被线粒体氧化呼吸链阻断剂myxothiazol阻断。结论：二氮嗪预处理对心肌细胞的保护作用与线粒体电子传递链氧化产生的氧自由基有关。     

低氧调节大鼠骨骼肌线粒体呼吸链复合体非协同性表达

目的观察慢性低氧时大鼠线粒体呼吸链复合体Ⅰ、Ⅱ、Ⅲ、Ⅴ蛋白表达变化并讨论其病理生理学意义。方法成年雄性Wistar大鼠随机分为慢性低氧(4500m,30d)组和对照组,取双侧腓肠肌,分离线粒体,用Clark氧电极法检测线粒体Ⅲ态呼吸(state3,ST3)、Ⅳ态呼吸(state4,ST4)和呼吸控制率(respiratory control ratio,RCR),用Western blot检测线粒体呼吸链复合体Ⅰ、Ⅱ、Ⅲ、Ⅴ蛋白的表达。结果慢性低氧组大鼠腓肠肌线粒体ST3和RCR显著低于对照组(P<0.05)。慢性低氧组大鼠腓肠肌线粒体复合体Ⅰ30×103亚基、复合体Ⅱ70×103亚基、复合体Ⅴα亚基蛋白表达量显著低对照组(P<0.05)。2组线粒体ST4和复合体Ⅲ核心亚基2蛋白表达无显著性差异(P>0.05)。结论低氧可调节大鼠骨骼机线粒体复合体蛋白非协同性表达,导致线粒体氧化磷酸化功能下降。     

SMO液对低温下肾皮质线粒体功能的保护作用

目的:探讨自制上海多器官保存液(SMO液)对肾皮质线粒体低温缺氧损伤的保护作用.方法:以HC-A液和UW液作对照,在犬肾低温保存各时间点,取皮质标本;通过电镜观察保存期间线粒体形态改变;差速离心法分离皮质线粒体,Clark氧电极法测定其活性,描记四态呼吸曲线,计算Ⅲ态、Ⅳ态耗氧速率、线粒体呼吸控制率(RCR)及磷氧比(P/O).结果:低温保存期间, HC-A组肾皮质线粒体水肿、空泡样变明显重于SMO组和UW组,而后两者改变相似;随着低温保存时间延长,实验组和对照组肾皮质线粒体Ⅲ态呼吸耗氧速率、RCR和P/O均明显降低;保存1～3 d HC-A组Ⅲ态呼吸耗氧速率显著低于SMO组(P<0.05);保存各时间点SMO组线粒体RCR和P/O显著高于HC-A组(P<0.05),与UW组无明显差异.结论:SMO液对肾皮质线粒体低温缺氧损伤的保护作用比HC-A液强,与UW液相似.     

脑缺血再灌流线粒体呼吸功能的变化

目的 观察鼠脑缺血和再灌流后脑细胞线粒体呼吸功能的改变。方法 采用沙鼠双侧颈总动脉夹闭30分钟，再开放造成脑缺血再灌流模型。测定脑缺血及再灌流后1h、3h、6h、24h和-48h线粒体呼吸Ⅲ、Ⅳ态及呼吸控制率(RCR)；测定线粒体呼吸链NADH—CoQ还原酶、琥珀酸脱氢酶活力。结果 脑缺血时线粒体呼吸控制率(RCR)和NADH—CoQ还原酶活力明显下降；再灌流初期代偿性增高，随着再灌流时间延长，线粒体呼吸Ⅳ态增加而呼吸Ⅲ态未能恢复，使RCR再次降低；NADH—CoQ还原酶、琥珀酸脱氢酶活力于再灌流后进一步下降，再灌流6小时后更为明显。结论 脑缺血可以造成线粒体功能损害，再灌流得到氧供使线粒体功能有所恢复，但再灌流可以造成二次打击，使线粒体无效耗氧增加，造成脑细胞的进一步损害。     

黄芪对培养心肌细胞过氧化氢损伤的保护作用

目的 :探讨中药黄芪 (Astragalus,AS)对培养的心肌细胞氧化性损伤的保护作用及其机制 .方法 :体外培养的新生大鼠心肌细胞 ,分为 3组 :①对照组 (Normal组 ) ;②H2 O2组 :H2 O2 (0 .2mmol/L)与心肌细胞共育 1h ;③黄芪 +H2 O2组 :黄芪处理心肌细胞 30min后 ,加入H2 O2 与心肌细胞共育1h .心肌细胞损伤以细胞线粒体活性和乳酸脱氢酶 (LDH)活性来表示 ,同时检测心肌细胞超氧化物歧化酶 (SOD)活性和丙二醛 (MDA)含量 ,均以比色法检测 .结果 :H2 O2 处理细胞后可使细胞LDH活性显著增高 (2 7± 3) μkat/L ,细胞线粒体活性明显下降 0 .36± 0 .0 4 ,SOD活性较正常细胞显著下降(2 31± 2 2 ) μkat/L ,MDA含量显著增高 (1 6± 3) μmol/L .黄芪处理细胞后可显著提高心肌细胞线粒体活性 0 .5 0± 0 .0 5 ,降低LDH活性 (1 6 .4± 1 .8) μkat/L ;并提高细胞抗氧化能力 ,表现为较H2 O2 处理组 ,SOD活性增高 (331± 4 0 ) μkat/L ,MDA含量下降 (1 0 .7± 2 .4 ) μmol/L .结论 :黄芪可对抗H2 O2 对心肌细胞的损伤 ,其机制与提高心肌细胞抗氧化损伤能力有关     

冷适应大鼠肝脏线粒体的呼吸功能及蛋白质合成

从正常和冷冻大鼠肝脏中分离出线粒体。用氧电极法测定鼠肝线粒体呼吸功能,包括ADP:O比值(P/O值),呼吸控制率(RCR),以[~3H]-Leu参入为指标,观察线粒体内蛋白质合成;以SDS-聚丙烯酰胺凝胶电泳方法观察线粒体内膜蛋白质的变化。结果表明:冷适应2、4周时P/O值和RCR降低,6周时两指标恢复至正常水平。冷适应1,2,4周时线粒体蛋白质合成明显降低。冷适应2,3周时线粒体内膜蛋白质电泳图谱呈现明显变化,冷适应组线粒体内膜有新的蛋白质条带出现。     

吉马酮调控JAK2/STAT3信号通路对卵巢癌SKOV3细胞增殖、迁移、侵袭的影响

背景 卵巢癌死亡率居于女性生殖系统恶性肿瘤的首位,仍需不断探索新的药物来改善其预后,近年来许多研究报道了吉马酮有抗肿瘤细胞作用.目的 探讨吉马酮对人卵巢癌SKOV3细胞增殖、迁移、侵袭能力的影响及其作用机制.方法 培养人卵巢癌SKOV3细胞,分别用0 μmol/L、70 μmol/L、140 μmol/L、210 μmol/L、280 μmol/L、350 μmol/L吉马酮作用于卵巢癌SKOV3细胞24 h、48 h、72 h,采用CCK-8法检测吉马酮对卵巢癌SKOV3细胞增殖能力的影响;采用细胞划痕实验及Transwell实验观察吉马酮对卵巢癌细胞迁移、侵袭能力的影响;Western blot法检测各组细胞JAK2/STAT3信号通路相关蛋白及肿瘤侵袭转移相关蛋白MMP2、MMP9的表达.结果 CCK-8实验结果提示吉马酮可以抑制人卵巢SKOV3细胞的增殖,并存在时间及浓度依赖性;细胞划痕实验显示0 μmol/L、140 μmol/L、280 μmol/L吉马酮组划痕愈合率分别为69.00％ ± 6.76％、40.33％ ± 5.21％、13.79％ ± 9.23％ (P<0.05);Transwell迁移实验显示0 μmol/L、140 μmol/L、280 μmol/L吉马酮组穿过小室细胞个数分别为466.5 ± 47.7、319.4 ± 41.2、149.7 ± 26.3(P<0.05);Transwell侵袭实验显示0 μmol/L、140 μmol/L、280 μmol/L吉马酮组穿过小室细胞个数分别为278.6 ± 71.8、161.0 ± 35.4、70.1 ± 24.9.与对照组相比,药物组对细胞迁移及侵袭有明显的抑制作用(P<0.05),并具有浓度依赖性;Western blot结果提示吉马酮可浓度依赖性下调p-JAK2、p-STAT3蛋白表达,降低p-JAK2/JAK2、p-STAT3/STAT3,还可浓度依赖性下调MMP2、MMP9蛋白表达(P<0.05).结论 吉马酮可明显抑制卵巢癌SKOV3细胞的增殖、迁移、侵袭能力,其机制可能与下调JAK2/STAT3信号通路活性、下调MMP2、MMP9蛋白表达有关.     

血清谷丙、谷草转氨酶连续监测法测定条件的选择

测定了SGPT、SGOT催化转氨基反应的底物L-丙氨酸、L-门冬氨酸和α-酮戊二酸的米氏常数(K_m)_8和α-酮戊二酸对SGPT、SGOT催化反应的抑制常数K_I;选择了以上三个底物和还原型辅酶Ⅰ的最适反应浓度;测定了指示酶——乳酸脱氢酶和苹果酸脱氢酶的最适用量以及SGPT、SGOT催化反应测定的延迟时间。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

CLINICAL SIGNIFICANCE OF SERUM CYTOKINES IL-1β,sIL-2R,IL-6,TNF-α,AND IFN-v IN ACUTE CORONARY SYNDROME

Objectives To explore serum cytokines levels (including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v) and their significance in patients with acute coronary syndrome (ACS) and the subsequent follow-ups, with attempt to estimate the role of various serum inflammatory markers in the diagnosis and assessment of ACS.Methods The study population include 40 patients with acute myocardial infarction (AMI), 40 patients with unstable angina pectoris (UAP), and 40 controls. Among the 80 patients, 60 patients attended a follow up 4 months later. Serum inflammatory markers including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v were measured by enzyme linked immunosorbent assay.Results Serum IL- 1 β, sIL-2R, IL-6, TNF-α were significantly higher in AMI group or UAP group compared to the control group and became significantly lower 4 months later in the follow-up patients. Serum levels of IFN-v shows no significant difference between AMI group or UAP group and controls, also showing no significant change when measured in follow up patients. There was no correlation between serum creatine kinase-MB isoenzyme levels and serum inflammatory markers either in UAP or AMI group. Furthermore, when divided into two subgroups using Wagner's QRS scoring system in the AMI group, there is no difference of each serum inflammatory marker between ≤ 6 scores group and ＞ 6 scores group.Conclusion Serum levels of certain inflammatory markers may have some diagnostic value for ACS, and can be a useful marker reflecting disease stability.     

Possible Spread of adenovirus type 3 from poultry to humans ：indirect evidence from an outbreak in China

Objective:To explore the epidemiology and etiology for an outbreak of acute respiratory tract infection that occurred in one county of Jiangsu Province, China 2004. Methods: Only cases meeting the case definition were included in the study. We reviewed the medical records of the cases who were admitted to the local hospitals, interviewed cases by a standard questionnaire, and then described the epidemiotogic features and analyzed risk factors by means of a case-control study. We collected pharyngeal swab specimens and sent them to different laboratories for isolation and culture. The laboratory used different detection methods such as DIP, PCR, electron microscope examination and microneutralization assay, to identify and then type the positive specimens. Results:A total of 871 cases were reported during the period from April 18 to July 4,2004. The distribution of onset times presented two peaks, one in late May and another in middle June. The epidemic occurred mainly in the elementary and junior high schools in ten townships of one county, and the mean age of the cases was 12 years (range 7 months to 18 years). The course of the disease was acute, and was characterized by fever accompanied with sore throat and tonsillitis. The WBC count of cases was normal or elevated. The mean duration of illness was 5 days (range 2 to 12 days). No fatalities from illness were reported. A case-control study indicated that the possible risk factors were close contact with a case and/or poultry before onset and sharing of towels among members of the family. The typical CPE was observed through inoculating pharyngeal swab specimens into the HEP-2 cell cultures in different laboratories. An infection of adenovirus type 3 was verified by detecting positive specimens in different methods. Conclusion:This investigation demonstrated that the acute respiratory infection in cases was caused by adenovirus type 3. Cases occurred in over 70 schools in ten townships in 2004, and the route of transmission was possibly close contact with cases or droplet transmission.